Question 1

Design your own PCR programme using the following primers to amplify a DNA fragment of 1.8 Kb. The reaction is set up in the evening and will be collected the following day.

Answer

Primers

Primer A GTACTTGACTGGCATGAC Annealing Temp = 49C

Primer B TAGGTAGCAATCGTGAGC Annealing Temp = 49C

Step 1 95C for 5 min.

Step 2 95C for 1 min.

Step 3 49C for 1 min.

Step 4 72C for 2 min.

Step 5 72C for 10 min.

Step 6 7C for overnight.

Step 7 7C for min.

Repeat Steps 2 - 4 for 25-30 cycles.

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